Writing and erasing O-GlcNAc on target proteins in the brain

在大脑中的目标蛋白上写入和擦除 O-GlcNAc

• 批准号: 10637668
• 负责人: Christina Woo
• 金额: $ 175.63万
• 依托单位: HARVARD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-15 至 2026-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10637668
• 关键词: Acceleration; Affect; Alzheimer disease prevention; Alzheimer' s Disease; Alzheimer' s disease brain; Alzheimer' s disease model; Alzheimer' s disease risk; Alzheimer' s disease therapy; Binding; Biological Assay; Biological Models; Biological Process; Brain; Brain region; Cells; Chemicals; Circadian Rhythms; Clinic; Clinical Trials; Complement; Cytoplasm; Cytoplasmic Protein; Data Set; Defect; Distress; Drosophila genus; Drosophila inturned protein; Enzymes; Evaluation; Event; Genes; Genetic study; Glucosamine; Glycoproteins; Goals; Homer protein; In Vitro; Intervention; Investigation; Joints; Knock-in; Letters; Libraries; Link; Maps; Measurement; Measures; Methods; Mitochondria; Mitochondrial Proteins; Modality; Modeling; Modification; Molecular; Neurodegenerative Disorders; Neurons; Nuclear; Nuclear Proteins; Nutrient; Nutrient availability; O-GlcNAc transferase; Organism; Outcome; Parkinson Disease; Pathogenesis; Pathology; Phenotype; Physiological; Physiological Processes; Play; Post-Translational Protein Processing; Prevention; Protein Engineering; Proteins; Proteome; Public Health; Regulation; Research Personnel; Role; Site; Sleep; Sleep Wake Cycle; Sleep disturbances; Symptoms; System; Tauopathies; Writing; behavioral study; cell type; circadian; disease phenotype; drug repurposing; flexibility; fly; glycoproteomics; in vitro Assay; in vivo; inhibitor; innovation; insight; knock-down; nanobodies; novel; peptide O-linked N-acetylglucosamine-beta-N-acetylglucosaminidase; prevent; remediation; sensor; sleep abnormalities; sleep regulation; small molecule; small molecule libraries; sugar; supportive environment; targeted treatment; tau Proteins; therapeutic target; tool

WRITING AND ERASING O-GLCNAC ON TARGET PROTEINS IN THE BRAIN PROJECT SUMMARY O-Linked N-acetyl glucosamine (O-GlcNAc) is a nutrient sensor that dynamically modifies nuclear, cytoplasmic, and mitochondrial proteins. Dysregulation of O-GlcNAc has been linked to disruptions in sleep and circadian rhythm and several neurodegenerative diseases, including Alzheimer’s Disease (AD). While sleep and circadian rhythm defects are distressing symptoms of AD and other tauopathies, sleep disturbance may be a major risk factor for AD and is thought to accelerate its pathology. Extensive studies on the association of O-GlcNAc to AD have led to the first clinical trials targeting O-GlcNAc for therapy. However, these inhibitors globally alter the O- GlcNAc proteome, where a more targeted strategy may provide greater benefit. A systematic investigation of the connection between the O-GlcNAc modification and sleep regulation and AD pathogenesis would significantly impact the discovery of novel mechanisms to provide new avenues for targeted prevention and therapy. O-GlcNAc is regulated by nutrient availability and the complementary activity of two enzymes: O-GlcNAc transferase (OGT) writes the modification and O-GlcNAcase (OGA) erases it from proteins. Recently, innovations in protein engineering and gene editing tools developed by the co-investigators have provided access to precise tuning of O-GlcNAc on specific neurons and desired target proteins in the brain of Drosophila model systems of sleep and AD. Here, we will capitalize on the joint expertise in the Woo Lab and Walker Lab to facilitate the first systematic study to measure, map, and manipulate O-GlcNAc from desired target proteins and in specific neurons in the brain to yield crucial insights to the pathogenesis of AD and novel chemical strategies for remediation. To meet this goal, we will take a three-pronged approach. We will first systematically examine the relationship between O-GlcNAc in specific neurons of Drosophila models of sleep and AD pathogenesis to identify the brain regions that are most dependent on O-GlcNAc regulation. Second, we will use a targeted writer and eraser of O-GlcNAc, developed through protein engineering, to systematically examine the role of O-GlcNAc on selected target proteins in the brain to identify drivers and potential targets for alleviating sleep disruptions and AD pathogenesis. Third, we will pursue the discovery of small molecules that selectively write and erase O-GlcNAc in vitro and in vivo, which will complement our protein engineering approaches and provide targeted alternatives to global inhibitors that are under evaluation for AD therapy in the clinic. The successful outcome of this proposal will afford validated Drosophila models of sleep and AD with neuron-specific manipulation of OGT and OGA or specific target proteins using target writers and erasers of O-GlcNAc, with associated maps of O-GlcNAc proteins and sites, in addition to new and selective small molecule editors of O- GlcNAc to enable more targeted therapeutic approaches in the long-term. Additionally, the systematic methods and tools to connect physiological measurements to molecular function developed here will be translatable to the study of the connection of O-GlcNAc to other neurodegenerative diseases and beyond.

O-GLCNAC在脑内靶蛋白上的写入和擦除 项目摘要 O-连接的N-乙酰葡糖胺（O-GlcNAc）是一种营养传感器，其动态修饰细胞核、细胞质、 和线粒体蛋白质。O-GlcNAc的失调与睡眠和昼夜节律的破坏有关 心律失常和几种神经退行性疾病，包括阿尔茨海默病（AD）。在睡眠和昼夜节律 节律缺陷是AD和其他tau蛋白病的令人痛苦的症状，睡眠障碍可能是主要风险 AD的因子，并被认为加速其病理。关于O-GlcNAc与AD相关性的广泛研究 已经导致了第一个针对O-GlcNAc治疗的临床试验。然而，这些抑制剂在全球范围内改变了O- GlcNAc蛋白质组，其中更有针对性的策略可能提供更大的益处。系统地调查了 O-GlcNAc修饰与睡眠调节和AD发病机制之间的联系将显着 影响新机制的发现，为靶向预防和治疗提供新途径。 O-GlcNAc受营养素可用性和两种酶的互补活性调节： 转移酶（OGT）写入修饰，而O-GlcNAcase（OGA）将其从蛋白质中删除。最近， 共同研究人员开发的蛋白质工程和基因编辑工具的创新提供了 获得O-GlcNAc对果蝇脑中特定神经元和所需靶蛋白的精确调节 睡眠和AD的模型系统。在这里，我们将利用Woo实验室和步行者实验室的联合专业知识 促进第一次系统性研究，以测量，映射和操纵来自所需靶蛋白的O-GlcNAc 和大脑中特定神经元的研究，以获得对AD发病机制和新化学物质的重要见解。 补救策略。为了实现这一目标，我们将采取三管齐下的办法。我们将首先系统地 研究睡眠和AD果蝇模型的特定神经元中O-GlcNAc之间的关系 发病机制，以确定最依赖于O-GlcNAc调节的脑区域。二是 使用通过蛋白质工程开发的O-GlcNAc的靶向写入器和擦除器， O-GlcNAc对大脑中选定的靶蛋白的作用，以确定缓解的驱动因素和潜在靶点 睡眠障碍和AD发病机制。第三，我们将致力于发现小分子， 在体外和体内写入和擦除O-GlcNAc，这将补充我们的蛋白质工程方法， 为临床上正在进行AD治疗评估的全局抑制剂提供靶向替代品。的 这一建议的成功结果将提供有效的睡眠和AD果蝇模型， 使用O-GlcNAc的靶标写入器和擦除器操纵OGT和OGA或特异性靶蛋白， O-GlcNAc蛋白和位点的相关图谱，以及新的和选择性的O-GlcNAc小分子编辑器。 GlcNAc，以实现更有针对性的长期治疗方法。此外，系统方法 这里开发的将生理测量与分子功能联系起来的工具， 研究O-GlcNAc与其他神经退行性疾病及其他疾病的联系。